Compositions and methods relating to solenopsins and their uses in treating neurological disorders and enhancing cognitive and physical performance

ABSTRACT

Provided herein are piperidine alkaloids and uses thereof in neurological disorders and physical enhancement applications.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims benefit of U.S. Provisional Application No.60/806,887, filed Jul. 10, 2006, which is hereby incorporated herein byreference in its entirety.

BACKGROUND OF THE INVENTION

A number of diseases and disorders involving the central nervous systemare known, such as, for example, stroke and related ischemic diseases,spinal cord injuries, peripheral nerve injuries, traumatic braininjuries, retinal degeneration, epilepsy (such as generalized, partial,or refractory epilepsy), neuropsychiatric disorders, pain disorders andneurodegenerative disorders. Alzheimer's disease (AD) is most commonlyassociated with deposition of amyloid-β (Aβ) peptide in the brain.Oxidative stress and inflammation are important pathogenic mechanisms ofAlzheimer's disease (AD) and related disorders. The presence of Aβpeptide in the brain plays an important role in the development ofaggregates that may result in neuronal damage and dysfunction andmicroglial activation and neuropathological features of AD. Aβ (with orwithout tau/neurofibrillary tangles (NFTs)) perturbs cellular propertiesmainly by oxidant stress and inflammation, which overwhelms the cellularantioxidant/anti-inflammatory defense-mechanisms. Currently, the onlyavailable treatments for AD are acetylcholinesterase inhibitors whichhave limited capabilities of reducing the effects of AD. However,current data suggests that the acetylcholinesterase inhibitorsaccelerate the process of AD. Therefore, it is crucial for thedevelopment of new therapeutics to treat the disease for AD as well asother neurological diseases and disorders.

BRIEF SUMMARY OF THE INVENTION

In accordance with the purpose of this invention, as embodied andbroadly described herein, this invention relates to piperidine alkaloidsand uses thereof in neurological applications.

Additional advantages of the disclosed method and compositions will beset forth in part in the description which follows, and in part will beunderstood from the description, or may be learned by practice of thedisclosed method and compositions. The advantages of the disclosedmethod and compositions will be realized and attained by means of theelements and combinations particularly pointed out in the appendedclaims. It is to be understood that both the foregoing generaldescription and the following detailed description are exemplary andexplanatory only and are not restrictive of the invention as claimed.

DETAILED DESCRIPTION OF THE INVENTION

The disclosed method and compositions may be understood more readily byreference to the following detailed description of particularembodiments and the Example included therein and to the Figures andtheir previous and following description.

It has been discovered that piperidine alkaloids, such as Solenopsin A,found in fire ant venom, can be used to treat and/or prevent, forexample, neurological disease or dysfunction or for the enhancement ofneurological or cognitive function. Other uses are disclosed, apparentfrom the disclosure, and/or will be understood by those in the art.

Disclosed are materials, compositions, and components that can be usedfor, can be used in conjunction with, can be used in preparation for, orare products of the disclosed method and compositions. These and othermaterials are disclosed herein, and it is understood that whencombinations, subsets, interactions, groups, etc. of these materials aredisclosed that while specific reference of each various individual andcollective combinations and permutation of these compounds may not beexplicitly disclosed, each is specifically contemplated and describedherein. For example, if an alkaloid is disclosed and discussed and anumber of modifications that can be made to a number of moleculesincluding the alkaloid are discussed, each and every combination andpermutation of alkaloid and the modifications that are possible arespecifically contemplated unless specifically indicated to the contrary.Thus, if a class of molecules A, B, and C are disclosed as well as aclass of molecules D, E, and F and an example of a combination molecule,A-D is disclosed, then even if each is not individually recited, each isindividually and collectively contemplated. Thus, is this example, eachof the combinations A-E, A-F, B-D, B-E, B-F, C-D, C-E, and C-F arespecifically contemplated and should be considered disclosed fromdisclosure of A, B, and C; D, E, and F; and the example combination A-D.Likewise, any subset or combination of these is also specificallycontemplated and disclosed. Thus, for example, the sub-group of A-E,B-F, and C-E are specifically contemplated and should be considereddisclosed from disclosure of A, B, and C; D, E, and F; and the examplecombination A-D. This concept applies to all aspects of this applicationincluding, but not limited to, steps in methods of making and using thedisclosed compositions. Thus, if there are a variety of additional stepsthat can be performed it is understood that each of these additionalsteps can be performed with any specific embodiment or combination ofembodiments of the disclosed methods, and that each such combination isspecifically contemplated and should be considered disclosed.

A. DEFINITIONS

It must be noted that as used herein and in the appended claims, thesingular forms “a,” “an,” and “the” include plural reference unless thecontext clearly dictates otherwise. Thus, for example, reference to “analkaloid” includes a plurality of such alkaloids, reference to “thealkaloid” is a reference to one or more alkaloids and equivalentsthereof known to those skilled in the art, and so forth.

“Optional” or “optionally” means that the subsequently described event,circumstance, or material may or may not occur or be present, and thatthe description includes instances where the event, circumstance, ormaterial occurs or is present and instances where it does not occur oris not present.

Ranges may be expressed herein as from “about” one particular value,and/or to “about” another particular value. When such a range isexpressed, also specifically contemplated and considered disclosed isthe range from the one particular value and/or to the other particularvalue unless the context specifically indicates otherwise. Similarly,when values are expressed as approximations, by use of the antecedent“about,” it will be understood that the particular value forms another,specifically contemplated embodiment that should be considered disclosedunless the context specifically indicates otherwise. It will be furtherunderstood that the endpoints of each of the ranges are significant bothin relation to the other endpoint, and independently of the otherendpoint unless the context specifically indicates otherwise. Finally,it should be understood that all of the individual values and sub-rangesof values contained within an explicitly disclosed range are alsospecifically contemplated and should be considered disclosed unless thecontext specifically indicates otherwise. The foregoing appliesregardless of whether in particular cases some or all of theseembodiments are explicitly disclosed.

It is understood that the disclosed method and compositions are notlimited to the particular methodology, protocols, and reagents describedas these may vary. It is also to be understood that the terminology usedherein is for the purpose of describing particular embodiments only, andis not intended to limit the scope of the present invention which willbe limited only by the appended claims.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meanings as commonly understood by one of skill in the artto which the disclosed method and compositions belong. Although anymethods and materials similar or equivalent to those described hereincan be used in the practice or testing of the present method andcompositions, the particularly useful methods, devices, and materialsare as described. Publications cited herein and the material for whichthey are cited are hereby specifically incorporated by reference.Nothing herein is to be construed as an admission that the presentinvention is not entitled to antedate such disclosure by virtue of priorinvention. No admission is made that any reference constitutes priorart. The discussion of references states what their authors assert, andapplicants reserve the right to challenge the accuracy and pertinency ofthe cited documents. It will be clearly understood that, although anumber of publications are referred to herein, such reference does notconstitute an admission that any of these documents forms part of thecommon general knowledge in the art.

Throughout the description and claims of this specification, the word“comprise” and variations of the word, such as “comprising” and“comprises,” means “including but not limited to,” and is not intendedto exclude, for example, other additives, components, integers or steps.

As used herein, “subject” can be a vertebrate, more specifically amammal (e.g., a human, horse, pig, rabbit, dog, sheep, goat, non-humanprimate, cow, cat, guinea pig or rodent), a fish, a bird or a reptile oran amphibian. The term does not denote a particular age or sex. Thus,adult and newborn subjects, as well as fetuses, whether male or female,are intended to be covered. The term “patient” refers to a subjectafflicted with a disease or disorder includes human and veterinarysubjects.

By “treatment” is meant the medical management of a patient with theintent to cure, ameliorate, stabilize, or prevent a disease,pathological condition, or disorder. This term includes activetreatment, that is, treatment directed specifically toward theimprovement of a disease, pathological condition, or disorder, and alsoincludes causal treatment, that is, treatment directed toward removal ofthe cause of the associated disease, pathological condition, ordisorder. In addition, this term includes palliative treatment, that is,treatment designed for the relief of symptoms rather than the curing ofthe disease, pathological condition, or disorder; preventativetreatment, that is, treatment directed to minimizing or partially orcompletely inhibiting the development of the associated disease,pathological condition, or disorder; and supportive treatment, that is,treatment employed to supplement another specific therapy directedtoward the improvement of the associated disease, pathologicalcondition, or disorder.

The term “therapeutically effective” means that the amount of thecomposition used is of sufficient quantity to ameliorate one or morecauses or symptoms of a disease or disorder. Such amelioration onlyrequires a reduction or alteration, not necessarily elimination. As usedherein, the term “substituted” is contemplated to include allpermissible substituents of organic compounds. In a broad aspect, thepermissible substituents include acyclic and cyclic, branched andunbranched, carbocyclic and heterocyclic, and aromatic and nonaromaticsubstituents of organic compounds. Illustrative substituents include,for example, those described below. The permissible substituents can beone or more and the same or different for appropriate organic compounds.For purposes of this disclosure, the heteroatoms, such as nitrogen, canhave hydrogen substituents and/or any permissible substituents oforganic compounds described herein which satisfy the valencies of theheteroatoms. This disclosure is not intended to be limited in any mannerby the permissible substituents of organic compounds. Also, the terms“substitution” or “substituted with” include the implicit proviso thatsuch substitution is in accordance with permitted valence of thesubstituted atom and the substituent, and that the substitution resultsin a stable compound, e.g., a compound that does not spontaneouslyundergo transformation such as by rearrangement, cyclization,elimination, etc.

“A¹,” “A²,” “A³,” and “A⁴” are used herein as generic symbols torepresent various specific substituents. These symbols can be anysubstituent, not limited to those disclosed herein, and when they aredefined to be certain substituents in one instance, they can, in anotherinstance, be defined as some other substituents.

The term “alkyl” as used herein is a branched or unbranched saturatedhydrocarbon group of 1 to 30 carbon atoms, such as methyl, ethyl,n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, pentyl, hexyl, heptyl,octyl, nonyl, decyl, dodecyl, tetradecyl, hexadecyl, eicosyl,tetracosyl, and the like. The alkyl group can also be substituted orunsubstituted. The alkyl group can be substituted with one or moregroups including, but not limited to, alkyl, halogenated alkyl, alkoxy,alkenyl, alkynyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid,ester, ether, halide, hydroxy, ketone, nitro, silyl, sulfo-oxo,sulfonyl, sulfone, sulfoxide, or thiol, as described below. The termalkyl also includes “cycloalkyl,” which is a cyclic alkyl group such ascyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like.

The term “alkoxy” as used herein is an alkyl group bound through asingle, terminal ether linkage; that is, an “alkoxy” group can bedefined as —OA¹ where A¹ is alkyl as defined above.

The term “alkenyl” as used herein is a hydrocarbon group of from 2 to 30carbon atoms with a structural formula containing at least onecarbon-carbon double bond. Asymmetric structures such as (A¹A²)C═C(A³A⁴)are intended to include both the E and Z isomers. This may be presumedin structural formulae herein wherein an asymmetric alkene is present,or it may be explicitly indicated by the bond symbol C═C. The alkenylgroup can be substituted with one or more groups including, but notlimited to, alkyl, halogenated alkyl, alkoxy, alkenyl, alkynyl, aryl,heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide,hydroxy, ketone, nitro, silyl, sulfo-oxo, sulfonyl, sulfone, sulfoxide,or thiol, as described below. The term alkenyl also includes“cycloalkenyl,” which is a cyclic alkenyl group such as cyclobutenyl,cyclopentenyl, cyclohexenyl, cycloheptenyl, and the like.

The term “alkynyl” as used herein is a hydrocarbon group of 2 to 30carbon atoms with a structural formula containing at least onecarbon-carbon triple bond. The alkynyl group can be substituted with oneor more groups including, but not limited to, alkyl, halogenated alkyl,alkoxy, alkenyl, alkynyl, aryl, heteroaryl, aldehyde, amino, carboxylicacid, ester, ether, halide, hydroxy, ketone, nitro, silyl, sulfo-oxo,sulfonyl, sulfone, sulfoxide, or thiol, as described below. The termalkyl also includes “cycloalkynyl,” which is a cyclic alkyl group suchas cyclobutynyl, cyclopentynyl, cyclohexynyl, cycloheptynyl, and thelike.

The term “aliphatic” is used herein to refer to an alkyl, alkenyl, oralkynyl group as defined herein.

The term “aryl” as used herein is a group that contains any carbon-basedaromatic group including, but not limited to, benzene, naphthalene,phenyl, biphenyl, phenoxybenzene, and the like. The term “aryl” alsoincludes “heteroaryl,” which is defined as a group that contains anaromatic group that has at least one heteroatom incorporated within thering of the aromatic group. Examples of heteroatoms include, but are notlimited to, nitrogen, oxygen, sulfur, and phosphorus. Likewise, the term“non-heteroaryl,” which is also included in the term “aryl,” defines agroup that contains an aromatic group that does not contain aheteroatom. The aryl group can be substituted or unsubstituted. The arylgroup can be substituted with one or more groups including, but notlimited to, alkyl, halogenated alkyl, alkoxy, alkenyl, alkynyl, aryl,heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide,hydroxy, ketone, nitro, silyl, sulfo-oxo, sulfonyl, sulfone, sulfoxide,or thiol as described herein. The term “biaryl” is a specific type ofaryl group and is included in the definition of aryl. Biaryl refers totwo aryl groups that are bound together via a fused ring structure, asin naphthalene, or are attached via one or more carbon-carbon bonds, asin biphenyl.

The term “aldehyde” as used herein is represented by the formula —C(O)H.Throughout this specification “C(O)” is a short hand notation for C═O.

The terms “amine” or “amino” as used herein are represented by theformula NA¹A²A³, where A¹, A², and A³ can be, independently, hydrogen,an alkyl, halogenated alkyl, alkenyl, alkynyl, aryl, heteroaryl,cycloalkyl, cycloalkenyl, heterocycloalkyl, or heterocycloalkenyl groupdescribed above.

The term “carboxylic acid” as used herein is represented by the formula—C(O)OH. A “carboxylate” as used herein is represented by the formula—C(O)O—.

The term “ester” as used herein is represented by the formula —OC(O)A¹or —C(O)OA¹, where A¹ can be an alkyl, halogenated alkyl, alkenyl,alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl,or heterocycloalkenyl group described above.

The term “ether” as used herein is represented by the formula A¹OA²,where A¹ and A² can be, independently, an alkyl, halogenated alkyl,alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl,heterocycloalkyl, or heterocycloalkenyl group described above.

The term “ketone” as used herein is represented by the formula A¹C(O)A²,where A¹ and A² can be, independently, an alkyl, halogenated alkyl,alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl,heterocycloalkyl, or heterocycloalkenyl group described above.

The term “halide” as used herein refers to the halogens fluorine,chlorine, bromine, and iodine.

The term “hydroxyl” as used herein is represented by the formula —OH.

The term “nitro” as used herein is represented by the formula —NO₂.

The term “silyl” as used herein is represented by the formula —SiA¹A²A³,where A¹, A², and A³ can be, independently, hydrogen, alkyl, halogenatedalkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl,cycloalkenyl, heterocycloalkyl, or heterocycloalkenyl group describedabove.

The term “sulfo-oxo” as used herein is represented by the formulas—S(O)A¹, —S(O)₂A¹, —OS(O)₂A¹, or —OS(O)₂OA¹, where A¹ can be hydrogen,an alkyl, halogenated alkyl, alkenyl, alkynyl, aryl, heteroaryl,cycloalkyl, cycloalkenyl, heterocycloalkyl, or heterocycloalkenyl groupdescribed above. Throughout this specification “S(O)” is a short handnotation for S═O

The term “sulfonyl” is used herein to refer to the sulfo-oxo grouprepresented by the formula —S(O)₂A¹, where A¹ can be hydrogen, an alkyl,halogenated alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl,cycloalkenyl, heterocycloalkyl, or heterocycloalkenyl group describedabove.

The term “sulfonylamino” or “sulfonamide” as used herein is representedby the formula —S(O)₂NH—.

The term “sulfone” as used herein is represented by the formulaA¹S(O)₂A², where A¹ and A² can be, independently, an alkyl, halogenatedalkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl,heterocycloalkyl, or heterocycloalkenyl group described above.

The term “sulfoxide” as used herein is represented by the formulaA¹S(O)A², where A¹ and A² can be, independently, an alkyl, halogenatedalkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl,heterocycloalkyl, or heterocycloalkenyl group described above.

The term “thiol” as used herein is represented by the formula —SH.

“R¹,” “R²,” “R³,” “Rn,” where n is an integer, as used herein can,independently, possess one or more of the groups listed above. Forexample, if R¹ is a straight chain alkyl group, one of the hydrogenatoms of the alkyl group can optionally be substituted with a hydroxylgroup, an alkoxy group, an alkyl group, a halide, and the like.Depending upon the groups that are selected, a first group can beincorporated within second group or, alternatively, the first group canbe pendant (i.e., attached) to the second group. For example, with thephrase “an alkyl group comprising an amino group,” the amino group canbe incorporated within the backbone of the alkyl group. Alternatively,the amino group can be attached to the backbone of the alkyl group. Thenature of the group(s) that is (are) selected will determine if thefirst group is embedded or attached to the second group.

Unless stated to the contrary, a formula with chemical bonds shown onlyas solid lines and not as wedges or dashed lines contemplates eachpossible isomer, e.g., each enantiomer and diastereomer, and a mixtureof isomers, such as a racemic or scalemic mixture.

B. COMPOSITIONS

Provided herein are piperidine alkaloids and uses thereof inneurological applications. These piperidine alkaloids can be derivedfrom venom, such as fire ant venom, or they can be synthesized asdisclosed herein.

1. Fire Ant Venom

Solenopsins are piperidine alkaloids derived from the venom of the redfire ant Solenopsis invicta. The term “fire ant” and “Solenopsisinvicta” are used interchangably to describe the common red fire ant,originating in South America, but now commonly found in 11 southeasternstates of the United States, parts of the southwestern United States,California, and Puerto Rico. The term fire ant may also be used todescribe black fire ants and other hybrid fire ants or other ants whichproduce solenopsin venom.

The venom of this insect consists of 95% alkaloids and the remaindercontains solubilized proteins, amino acids and enzymes includinghyaluronidase and phospholipase. Among the piperidine alkaloids in thevenom, the two major components are Solenopsin A, a trans-2-methyl6-n-undecylpiperidine and Solenopsin B, a 2,6-trans-dialkyl-piperidine.

The use of solenopsins for the elimination of ticks, fleas or otherparasitic infections in dogs and cats has been disclosed by Rehmert etal. in U.S. Pat. Nos. 4,910,209, 5,075,320, and 5,098,914, which arehereby incorporated by reference herein in their entirety for theseteachings. In this approach, the solenopsins were administered from thewhole body extract of the insect or from an oral dosage form containingmore highly purified material. The administration of these drugs over aperiod of one to eleven days with regular booster dosages disseminatesthe alkaloid composition through the blood and tissue fluids of thetreated animals and eliminates fluid-feeding parasites.

2. Piperidine Alkaloids

Piperidine alkaloids can be used in the disclosed compositions andmethods. Piperidine alkaloids, such as Solenopsin A, can also besynthetically produced. Unlike the whole body extract, it does notrequire refrigeration. However, higher numbers of units of the syntheticversion are required for effective treatment. For example, 1,500 unitsof synthetically produced Solenopsin A are equivalent to 250 units ofthe whole body extracts.

The stings of Red Imported Fire Ants (Solenopsis invicta Buren) containhigh alkaloid content in the venom, which exhibits potent necrotoxicactivity. The major components of the venom have various cis and transisomers of 2-methyl-6-n-alkyl piperidines, with the trans isomerpredominating.

The chemical structures of the piperidine alkaloid, Solenopsin A, areshown below.

wherein R¹ is a C11 alkyl group (i.e., undecyl group). However, othercompounds that can be present in venom in varying amounts, and are alsosuitable for the uses and compositions disclosed herein, include, butare not limited to,

-   I. cis-2-methyl-6-undecylpiperidine-   II. trans-2-methyl-6-undecylpiperidine-   III. cis-2-methyl-6-tridecylpiperidine-   IV. trans-2-methyl-6-tridecylpiperidine-   V. cis-2-methyl-6-pentadecylpiperidine-   VI. trans-2-methyl-6-pentadecylpiperidine-   VII. cis-2-methyl-6-(cis-4-tridecen-1-yl)piperidine-   VIII. trans-2-methyl-6-(cis-4-tridecen-1-yl)piperidine-   IX. cis-2-methyl-6-(cis-4-pentadecen-1-yl)piperidine-   X. trans-2-methyl-6-(cis-4-pentadecen-1-yl)piperidine

Other examples of piperidine alkaloids that can be used in the disclosedcompositions and methods can have the following general formula.

wherein R¹ is a short chain (i.e., 1-6 carbon atoms) substituted orunsubstituted aliphatic group, and R² is a long chain substituted orunsubstituted aliphatic group having from 7 to 30 carbon atoms. In manyspecific examples, R¹ is methyl, ethyl, or propyl, and R¹ is a alkyl oralkenyl group having from 10 to 20 carbons (i.e., 10, 11, 12, 13, 14,15, 16, 17, 18, 19, or 20 carbons, including mixtures thereof).

Also contemplated herein are pharmaceutically acceptable salts of thedisclosed piperidine alkaloids. Such salts can be prepared by treatingthe piperidine alkaloids with an appropriate amount of apharmaceutically acceptable acid such as, for example, HCl, H₂SO₄, orH₃PO₄, to produce the cationic salt. In one example, the compound can beprotonated with tartaric acid or acetic acid to produce the tartarate oracetate salt, respectively. In another example, the reaction of thecompound with the acid is conducted in water, alone or in combinationwith an inert, water-miscible organic solvent, at a temperature of fromabout 0° C. to about 100° C., such as at room temperature. In certainsituations, where applicable, the molar ratio of the disclosed compoundsto base is chosen to provide the ratio desired for any particular salts.

3. Combinations

The methods and compositions disclosed herein can be used in combinationwith various compositions. For example, the following drugs and classesof drugs can be used for treatment of neurological disorders: opioidsand opioid peptides, morphine, hydroxymorphine, fentanyl, oxycodone,codeine; capsaicin; as well as antiepileptic drugs as a class includingbut not limited to carbamazepine, primidone, gabapentin, pregabalin,diazepam, felbamate, fluorofelbamate, lamotrigine, lacosamide,levetiracetam, phenobarbital, phenyloin, fos-phenyloin, topiramate,valproate, vigabatrin, zonisamide, oxcarbazepine, nonsteroidalanti-inflammatory drugs (NSAIDs), local anesthetics (such as lidocaine),glutamate receptor antagonists, NMDA antagonists, alpha-adrenoceptoragonists and antagonists, adenosine, cannabinoids, NK-1 antagonist(CI-1021), antidepressants (amitriptyline, desipramine, imipramine, forexample), analogs and derivatives of galanin, somatostatin, delta-sleepinducing peptide, enkephalins, oxytocin. cholecystikinin, calcitonin,cortistatin, nociceptin and other neuropeptide-based therapeutics, andpluronic P85 block copolymer.

4. Pharmaceutical Compositions

The present compounds or their derivatives, including prodrug forms ofthese agents, can be provided in the form of pharmaceutically acceptablesalts. As used herein, the term pharmaceutically acceptable salts orcomplexes refers to appropriate salts or complexes of the activecompounds as disclosed herein which retain the desired biologicalactivity of the parent compound and exhibit limited toxicologicaleffects to normal cells. Nonlimiting examples of such salts are (a) acidaddition salts formed with inorganic acids (for example, hydrochloricacid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, andthe like), and salts formed with organic acids such as acetic acid,oxalic acid, tartaric acid, succinic acid, malic acid, ascorbic acid,benzoic acid, tannic acid, pamoic acid, alginic acid, and polyglutamicacid, among others; (b) base addition salts formed with metal cationssuch as zinc, calcium, sodium, potassium, magnesium, manganese and thelike, among numerous others, many of which may increase the watersolubility of the compounds in final pharmaceutical dosage form.

Modifications of the active compound can affect the solubility,bioavailability and rate of metabolism of the active species, thusproviding control over the delivery of the active species. The term“water soluble salt form” or “salt form” is used to describe forms ofcompounds as disclosed herein which are in their water soluble saltform. Salt forms of compounds as disclosed herein include any salt whichretains the desired biological effects. Nonlimiting examples of suchsalts are acid addition salts formed with inorganic acids such ashydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid,nitric acid and the like, salts formed with organic acids such as aceticacid, oxalic acid, tartaric acid, succinic acid, malic acid, ascorbicacid, benzoic acid, tannic acid, pamoic acid, alginic acid, glutamicacid, naphthalenesulfonic acids, naphthalenedisulfonic acids andglacturonic acid, among numerous others. Salts may be formed byneutralizing the nitrogen on the piperidine ring with the resultingsalts exhibiting substantially greater solubility or derliverability ofthe instant compounds. These may also affect the bioavailability andrate of metabolism or stability of the compounds as disclosed herein.

The disclosed compositions can be used therapeutically in combinationwith a pharmaceutically acceptable carrier. Thus, a piperidine alkaloidsdisclosed herein can be formulated in admixture with a pharmaceuticallyacceptable carrier, excipient or additive. By “pharmaceuticallyacceptable” is meant a material that is not biologically or otherwiseundesirable, i.e., the material may be administered to a subject, alongwith the disclosed piperidines, without causing any undesirablebiological effects or interacting in a deleterious manner with any ofthe other components of the pharmaceutical composition in which it iscontained. The carrier would naturally be selected to minimize anydegradation of the active ingredient and to minimize any adverse sideeffects in the subject, as would be well known to one of skill in theart.

Suitable carriers and their formulations are described in Remington: TheScience and Practice of Pharmacy (19th ed.) ed. A. R. Gennaro, MackPublishing Company, Easton, Pa. 1995. Typically, an appropriate amountof a pharmaceutically-acceptable salt is used in the formulation torender the formulation isotonic. Examples of thepharmaceutically-acceptable carrier include, but are not limited to,saline, Ringer's solution and dextrose solution. The pH of the solutionis preferably from about 5 to about 8, and more preferably from about 7to about 7.5. Further carriers include sustained release preparationssuch as semipermeable matrices of solid hydrophobic polymers containingthe antibody, which matrices are in the form of shaped articles, e.g.,films, liposomes or microparticles. It will be apparent to those personsskilled in the art that certain carriers may be more preferabledepending upon, for instance, the route of administration andconcentration of composition being administered.

Pharmaceutical carriers are known to those skilled in the art. Thesemost typically would be standard carriers for administration of drugs tohumans, including solutions such as sterile water, saline, and bufferedsolutions at physiological pH. The compositions can be administeredintramuscularly or subcutaneously. Other compounds will be administeredaccording to standard procedures used by those skilled in the art.

Pharmaceutical compositions may include carriers, thickeners, diluents,buffers, preservatives, surface active agents and the like in additionto the molecule of choice. Pharmaceutical compositions may also includeone or more active ingredients such as antimicrobial agents,antiinflammatory agents, anesthetics, and the like.

Preparations for parenteral administration include sterile aqueous ornon-aqueous solutions, suspensions, and emulsions. Examples ofnon-aqueous solvents are propylene glycol, polyethylene glycol,vegetable oils such as olive oil, and injectable organic esters such asethyl oleate. Aqueous carriers include water, alcoholic/aqueoussolutions, emulsions or suspensions, including saline and bufferedmedia. Parenteral vehicles include sodium chloride solution, Ringer'sdextrose, dextrose and sodium chloride, lactated Ringer's, or fixedoils. Intravenous vehicles include fluid and nutrient replenishers,electrolyte replenishers (such as those based on Ringer's dextrose), andthe like. Preservatives and other additives may also be present such as,for example, antimicrobials, anti-oxidants, chelating agents, and inertgases and the like.

Formulations for topical administration may include ointments, lotions,creams, gels, drops, suppositories, sprays, liquids and powders.Conventional pharmaceutical carriers, aqueous, powder or oily bases,thickeners and the like may be necessary or desirable.

Compositions for oral administration include powders or granules,suspensions or solutions in water or non-aqueous media, capsules,sachets, or tablets. Thickeners, flavorings, diluents, emulsifiers,dispersing aids or binders may be desirable.

Some of the compositions may potentially be administered as apharmaceutically acceptable acid- or base-addition salt, formed byreaction with inorganic acids such as hydrochloric acid, hydrobromicacid, perchloric acid, nitric acid, thiocyanic acid, sulfuric acid, andphosphoric acid, and organic acids such as formic acid, acetic acid,propionic acid, glycolic acid, lactic acid, pyruvic acid, oxalic acid,malonic acid, succinic acid, maleic acid, and fumaric acid, or byreaction with an inorganic base such as sodium hydroxide, ammoniumhydroxide, potassium hydroxide, and organic bases such as mono-, di-,trialkyl and aryl amines and substituted ethanolamines.

C. METHODS

As disclosed herein, piperidine alkaloids can be used for the treatmentof neurological disease or dysfunction or for the enhancement ofneurological or cognitive function. Other uses are disclosed, apparentfrom the disclosure, and/or will be understood by those in the art.

Without wishing to be bound by theory, Solenopsin can, in some aspects,have its effect on neurological or cognitive function by inhibitingphosphatidylinositol-3-kinase (PI3K) signaling. Beta secretase (BACE)expression results in increased intracellular Aβ 40-42 levels throughthe processing of amyloid precursor protein (APP). Thus, the hereindisclosed piperidine alkaloids can be used in combinations with otherPI3K inhibitors. In addition, treatment of a subject with the disclosedpiperidine alkaloids can be monitored by detecting PI3K activity intarget cells of the subject. This in turn can reduce APP processing inthe cell such as through the inhibition of BACE activity. A reduction inAPP processing can limit the production of Aβ peptides (1-40 and 1-42)and curb the extent of Alzheimer's disease (Aβ).

1. Neurological Disease

The methods and compositions disclosed herein can also be used in theprevention, amelioration, or treatment of a variety of diseases anddisorders involving the central nervous system, such as, for example,stroke and related ischemic diseases, spinal cord injuries, peripheralnerve injuries, traumatic brain injuries, retinal degeneration, epilepsy(such as generalized, partial, or refractory epilepsy), neuropsychiatricdisorders, and neurodegenerative disorders.

Neuropsychiatric disorders include, but are not limited to,schizophrenia, schizoaffective disorder, attention deficit disorder,dysthymic disorder, major depressive disorder, mania,obsessive-compulsive disorder, psychoactive substance use disorders,anxiety, panic disorder, as well as bipolar affective disorder, e.g.,severe bipolar affective (mood) disorder (BP-I), bipolar affective(mood) disorder with hypomania and major depression (BP-II). FurtherCNS-related disorders include, for example, those listed in the AmericanPsychiatric Association's Diagnostic and Statistical manual of MentalDisorders (DSM), the most current version of which is incorporatedherein by reference in its entirety.

Non-limiting examples of neurodegenerative disorders include Alexanderdisease, Alper's disease, Alzheimer disease, Amyotrophic LateralSclerosis (ALS, Lou Gehrig's disease), Ataxia telangiectasia, Battendisease (also known as Spielmeyer-Vogt-Sjogren-Batten disease), Canavandisease, Cockayne syndrome, Corticobasal degeneration, Creutzfeldt-Jakobdisease, Huntington disease, Kennedy's disease, Krabbe disease, Lewybody dementia, Machado-Joseph disease, Spinocerebellar ataxia type 3),Multiple sclerosis, Multiple System Atrophy, Parkinson disease,Pelizaeus-Merzbacher Disease, Pick's disease, Primary lateral sclerosis,Refsum's disease, Sandhoff disease, Schilder's disease,Spielmeyer-Vogt-Sjogren-Batten disease (also known as Batten disease),Spinocerebellar ataxia (multiple types with varying characteristics),Spinal muscular atrophy, Steele-Richardson-Olszewski disease, and Tabesdorsalis.

Alzheimer's disease is a progressive neurodegenerative disorder that ischaracterized by the formation of senile plaques and neurofibrillarytangles containing amyloid β (Aβ) peptide. These plaques are found inlimbic and association cortices of the brain. The hippocampus is part ofthe limbic system and plays an important role in learning and memory. Insubjects with Alzheimer's disease, accumulating plaques damage theneuronal architecture in limbic areas and eventually cripple the memoryprocess.

Approximately twenty million people worldwide suffer with dementia thatresults from Alzheimer's disease. The disease can be early onsetaffecting individuals as young as 30 years of age, or it can be familialor sporadic. Familial Alzheimer's disease was once thought to beinherited strictly as an autosomal dominant trait; however, this view ischanging as more genetic determinants are isolated. For example, somenormal allelic variants of apolipoprotein E (ApoE), which is found insenile plaques, can either protect against or increase the risk ofdeveloping the disease (Strittmatter et al. (1993) Proc Natl Acad Sci90:1977-1981).

Amyloid-β (Aβ) peptides are metabolites of the Alzheimer'sdisease-associated precursor protein, β-amyloid precursor protein (APP),and are believed to be the major pathological determinants ofAlzheimer's disease (AD). These peptides consist mainly of 40 to 42amino acids, Aβ1-40 (“Aβ40”) and Aβ1-42 (“Aβ42”), respectively. Aβ40 andAβ42 are generated by two enzymatic cleavages occurring close to theC-terminus of APP. The enzymes responsible for the cleavage, β-secretaseand γ-secretase, generate the - and C-termini of Aβ, respectively. Theamino terminus of Aβ is formed by β-secretase cleavage betweenmethionine residue 596 and aspartate residue 597 of APP (APP 695 isoformnumbering) (see, e.g., U.S. Pat. No. 6,440,698; and U.S. Pat. No.5,744,346).

γ-secretase activity cleaves at varying positions 38-, 40- or43-residues C-terminal of this β-secretase cleavage to release Aβpeptides (see, e.g., U.S. Patent Application 20020025540). The completemolecular identity of γ-secretase enzyme is still unknown. Presenilin 1,or the closely related presenilin 2, is needed for γ-secretase activity.γ-secretase activity is reduced 80% in cultured cells derived fromembryos genetically deleted for presenilin 1. All γ-secretase activityis lost in cells lacking both presenilin 1 and presenilin 2.Peptidomimetic inhibitors of γ-secretase activity can be crosslinked topresenilins 1 and 2, suggesting that these proteins are catalyticsubunits for the cleavage. However, γ-secretase activity isolated fromcells chromatographs as a large complex>1 M daltons. Genetic studieshave identified three more proteins required for γ-secretase activity;nicastrin, aph-1 and pen-1. (Francis et al., 2002, Developmental Cell3(1): 85-97; Steiner et al., 2002, J. Biol. Chemistry: 277(42):39062-39065; and Li et al., 2002, J. Neurochem. 82(6): 1540-1548).Accumulation of presenilin into high molecular weight complexes isaltered in cells lacking these proteins.

A third enzyme, α-secretase, cleaves the precursor protein between theβ- and γ-cleavage sites, thus precluding AP production and releasing anapproximately 3 kDa peptide known as P3, which is non-pathological. Bothβ- and α-secretase cleavage also result in soluble, secreted—terminalfragments of APP, known as sAPβ and sAPPα, respectively. The sAPPαfragment has been suggested to be neuroprotective.

In normal individuals, the Aβ peptide is found in two predominant forms,the majority Aβ-40 (also known as Aβ1-40) form and the minority A#42(also known as Aβ1-42) form, each having a distinct COOH-terminus. Themajor histological lesions of Aβ are neuritic plaques andneurofibrillary tangles occurring in affected brain regions. Neuriticplaques consist of Aβ peptides, primarily Aβ40 and Aβ42. Althoughhealthy neurons produce at least ten times more Aβ40 compared to Aβ42,plaques contain a larger proportion of the less soluble Aβ42. Patientswith the most common form of familial Alzheimer's disease show anincrease in the amount of the Aβ42 form. The Aβ40 form is not associatedwith early deposits of amyloid plaques. In contrast, the Aβ42 formaccumulates early and predominantly in the parenchymal plaques and thereis strong evidence that Aβ42 plays a major role in amyloid plaquedeposits in familial Alzheimer's disease patients (Roher et al., 1993,Proc. Natl. Acad. Sci. USA 90:10836; Iwatasubo, T., et al., 1994 Neuron13:45; Yamaguchi et al, 1995, Amyloid Int. J. Clin. Invest. 2:7-16; andMann et al., 1996 μm. J. Pathol. 148:1257).

Mutations in four genes are known to predispose an individual toAlzheimer's disease: ApoE, amyloid precursor protein (APP),presenilin-1, and presenilin-2 (Selkoe (1999) Nature 399:A23-A31). Thee4 allele of the ApoE gene confers increased risk for late onsetAlzheimer's disease. β-amyloid protein (Aβ) is the major component ofsenile plaques, and it is normally formed when β- and γ-secretase cleaveAPP. In Alzheimer's disease patients, large quantities of Aβ aregenerated and accumulate extracellularly in these neuropathologicalplaques. Efforts to understand the mechanism underlying Aβ depositionhave recently focused on the APP-cleaving secretase. In fact, two yeastaspartyl proteases have been shown to process APP in vitro (Zhang et al.(1997) Biochem Biophys Acta 1359:110-122). Evidence using peptidomimeticprobes further confirms that the secretase is an intramembrane-cleavingaspartyl protease (Wolfe et al. (1999) Biochemistry 38:4720-4727). Thepresenilin-1 gene is a candidate for the γ-secretase that cleaves theAPP carboxyl terminus. Several lines of evidence support the involvementof presenilins in the disease process. Presenilin can becoimmunoprecipitated with APP, and mutations in the presenilin genesincrease production of the 42-amino acid peptide form of Aβ. Thesemissense point mutations result in a particularly aggressive, earlyonset form of the disease (Haaas and DeStrooper (1999) Science286:916-919).

The proteases, BACE1 and BACE2 (β-site APP cleaving enzymes 1 and 2)which appear to be β-secretase, are potential therapeutic targetsbecause of their ability to cleave APP. Vassar et al. (1999; Science286:735-741) have found that BACE1 is an aspartyl protease withβ-secretase activity which cleaves APP to produce Aβ peptide in vitro.It is expressed at moderate levels across all brain regions and isconcentrated in neurons but not in glia. BACE2, which has 52% amino acididentity with BACE1, has been described by Saunders et al. (1999;Science 286:1255a). Whereas BACE1 maps to the long arm of chromosome 11,BACE2 maps to the Down syndrome region of chromosome 21 (Acquati et al.(2000) 468: 59-64; Saunders et al. supra). This location is significantbecause middle-aged Down syndrome patients have enhanced β-amyloiddeposits. Other members of the BACE family can also participate in thisAPP cleavage: the amino terminals of Aβ peptides appear to be cleavedheterogeneously indicating that there can be several β-secretaseinvolved in APP processing (Vassar (1999) Science 286:735-741).

Associations between Alzheimer's disease and many other genes andproteins have been reported. Fetal Alzheimer antigen (FALZ) andsynuclein a (SNCA) are found in brain plaques and tangles. Inheritanceof some gene polymorphisms is also linked to increased risk ofdeveloping the disease. For example, a polymorphism in the gene encodingβ2-macroglobulin, a protein that can act as a protease inhibitor, isassociated with increased risk for developing a late-onset form ofAlzheimer's disease.

One hundred years ago Alois Alzheimer described the major behavioral andneuropathological features of the neurodegenerative disorder bearing hisname. AD is characterized clinically/behaviorally by progressiveimpairment of memory and cognition. Neuropathological andneurobiological changes associated with this slow progression ofclinical symptoms include accumulation of amyloid plaques andneurofibrillary tangles (NFTs) (Gearing M. et al., The Consortium toEstablish a Registry for Alzheimer's Disease (CERAD). Part X.Neuropathology confirmation of the clinical diagnosis of Alzheimer'sdisease. Neurology. 1995; 45(3 Pt 1):461-466) gliosis (Unger J W.,Microscopy Res. Technique. 1998; 43:24-28), reduced dendritic plasticityrelative to normal aged (Buell S J. Coleman P D., Science. 1979;206(4420):854-856), Flood D G. et al., Brain Research. 1985;345(2):366-368, Flood D G., et al., Brain Research. 1987;402(2):205-216), and reduced density of neurons (Coleman P D. Flood DG., et al., Neurobiology of Aging. 1987; 8(6):521-545), Terry R D, etal., 1987; 21:530-539, West M J, et al., Lancet. 1994; 344:769-772) andsynapses (Scheff S W. et al., Neurobiology of Aging. 1990; 11(1):29-37).

Studies of altered gene expression in Alzheimer's disease brain tissuehave shown a general reduction of message level estimated at about 35%(Doebler J A, et al., Journal of Neuropathology & ExperimentalNeurology. 1987; 46(1):28-39), (Griffin W S, et al., Alzheimer Disease &Associated Disorders. 1990; 4(2):69-78), (Harrison P J, et al.,Psychological Medicine. 1991; 21:855-866). Against this background of ageneral reduction of mRNA, selected studies have demonstrated increasedas well as decreased expression of a wide variety of genes. Some geneclasses affected in Alzheimer's disease are expressed in a neuronspecific manner. These especially include decreased expression ofselected genes that are related to synaptic structure and function andthe neuronal cytoskeleton (Ginsberg S D. et al., Annals of Neurology.2000; 48(1):77-87), (Yao P, et al., Journal of Neuroscience. 1998;18(7):2399-2411). Other classes of genes whose expression is altered inAD include those related to the cell cycle (Arendt T., Neurobiology ofAging. 2000; 21(6):783-796), (Husseman J W., et al., Neurobiology ofAging. 2000; 21(6):815-828), (Nagy Z., et al., Neurobiology of Aging.2000; 21(6):761-769), (Vincent I, et al., J. Neurosci. 1997;17:3588-3598) and inflammatory/stress responses (for a review, see(Akiyama H., et al., Neurobiology of Aging. 2000; 21(3):383-421)). Thesegene classes are expressed in a variety of cell types that resideoutside the nervous system including leukocytes (Wakutani Y. et al.,Dementia. 1995; 6(6):301-305), monocytes (Jung S S. et al., Neurobiologyof Aging. 1999; 20(3):249-257), and epithelial cells (Schmitz A., etal., Histochemistry & Cell Biology. 2002; 117(2):171-180) as well asother cell types.

Multivariate analysis of profiles of expression of multiple geneproducts (messages) by single neurons or homogenates from postmortemhuman brain can be used to distinguish Alzheimer's disease from controlsamples (Cheetham J E., et al., Journal of Neuroscience Methods. 1997;77(1):43-48, Chow, N., et al., Proc. Natl. Acad. Sci. USA. 1998;95:9620-9625).

Symptoms of Aβ-related disorders are well known to those of skill in theart. For example, symptoms of Alzheimer's disease are well known in theart and can include, e.g., memory loss, mild cognitive impairment,cognitive decline, severe cognitive impairment and personality changesthat result in loss of functional ability, e.g., over the course of adecade. In debilitated states, patients usually exhibit severeimpairment, and retain only vegetative neurologic function. Symptoms ofAlzheimer's disease can also include certain art-known neuropathologicallesions, including intracellular neurofibrillary tangles andextracellular parenchymal and cerebrovascular amyloid.

Thus, provided is a method of treating or preventing Alzheimer's diseasein a subject comprising administering to the subject a therapeuticallyeffective amount of a composition comprising a piperidine alkaloid. Alsoprovided is a method of treating a subject at risk for Alzheimer'sdisease comprising administering to the subject a composition comprisinga piperidine alkaloid. As used herein, the terms “disorder” and“disease” are used interchangeably to refer to a condition in a subject.

As used herein, the term “Aβ-related disorder” or an “Aβ disorder” is adisease (e.g., Alzheimer's disease) or a condition (e.g., seniledementia) that involves an aberration or dysregulation of Aβ levels. AnAβ-related disorder includes, but is not limited to Alzheimer's disease,Down's syndrome and inclusion body myositis. Thus, the Aβ relateddisorder can be Alzheimer's disease. The progression of the Aβ relateddisorder can be slowed or reversed. Also provided is a method formodulating amyloid-β peptide (Aβ) levels exhibited by a cell or tissuecomprising contacting said cell or tissue with an amount of acomposition comprising a piperidine alkaloid sufficient to modulate saidAβ levels.

As used herein, a cell or tissue may include, but not be limited to: anexcitable cell, e.g., a sensory neuron, motomeuron, or interneuron; aglial cell; a primary culture of cells, e.g., a primary culture ofneuronal or glial cells; cell(s) derived from a neuronal or glial cellline; dissociated cell(s); whole cell(s) or intact cell(s);permeabilized cell(s); a broken cell preparation; an isolated and/orpurified cell preparation; a cellular extract or purified enzymepreparation; a tissue or organ, e.g., brain, brain structure, brainslice, spinal cord, spinal cord slice, central nervous system,peripheral nervous system, or nerve; tissue slices, and a whole animal.In certain embodiments, the brain structure is cerebral cortex, thehippocampus, or their anatomical and/or functional counterparts in othermammalian species. In certain embodiments, the cell or tissue is an N2acell, a primary neuronal culture or a hippocampal tissue explant.

2. Enhance Neurological Function and Physical Performance

The methods and compositions disclosed herein can also be used in theenhancement of neurological and/or cognitive function. Cognitiveenhancement may be defined for present purposes as a measurableimprovement in a cognitive ability of a mammalian subject. Methods andmeans of measuring cognitive abilities of experimental laboratoryanimals, such as rats, are well-known to those skilled in the art (e.g.,shuttle boxes, Morris-mazes, etc). Similarly, there are methods ofmeasuring the cognitive abilities of human subjects (e.g., as employedby Becker et al) known to, those skilled in the art. A number of testshave been used to investigate the cognitive abilities of Alzheimer'sDisease patients in clinical trials to assess the effectiveness of drugtherapies. Examples include the “Alzheimer Disease Assessment Scale”(ADAS-Cog) (Rosen et al, Am. J. Psychiatry 1984 141, 1356-1364) and the“Mini Mental State Examination” (MMSE) (Rosen et al, J. Psychiatric Res.1975 12,189-198). Cognitive enhancement is detected by a statisticallysignificant improvement (e.g. in the test group receiving the drugcompared to a control group, as measured by an appropriate statisticaltest (e.g. Student's T test).

The methods and compositions disclosed herein can also be used in theenhancement of physical performance. Physical performance enhancementmay be defined for present purposes as a measurable improvement in aphysical ability of a mammalian subject. Methods and means of measuringphysical abilities of experimental laboratory animals, such as rats, arewell-known to those skilled in the art (e.g., grip strength, treadmill,etc). Similarly, there are methods of measuring the physical abilitiesof human subjects known to those skilled in the art. A number of testshave been used to investigate the physical abilities of individuals.

3. Method of Making

The disclosed piperidine alkaloids can be found in or extracted from antvenom or can be produced synthetically.

i. Extracts

Ant venom extracts can be made from ant venom or whole ants. For wholebody extracts, insects can be ground to a fine texture, inserted intosoluble capsules as whole body extract along with an edible carriermaterial such as fish oil, and are kept frozen until administration. Thevenom can be kept refrigerated in order to maintain its effectiveness.Additionally, each insect can contain approximately one venom unit or 40nanoliters of the solenopsins, Solenopsin A and Solenopsin B.

ii. Chemical Synthesis

The present compounds can also be synthesized using general methodswhich are well known in the art. For examples, Wilkinson et al. reporteda method for the enantioselective synthesis of 2,6-disubstitutedpiperidine alkaloids in Org. Lett. 2(2)155-8, 2000. Monfray et al. alsoreported a method for the synthesis of cis-2,6-disubstituted and2,4,6-trisubstituted piperidine alkaloids (Tetrahedron Asymm.16(5):1025-34, 2005). Hiroki et al. reported a method for thepreparation of trans-2,6-disubstituted piperidine alkaloids (Org.Biomol. Chem. 4:1587-95, 2006). Still further examples are reported byBeak and Lee, who use Boc (tert-butoxycarbonyl) protected cyclic aminesand convert them into 2,6-disubstituted piperidines (J. Org. Chem.58:1109-1117, 1993). Further, Yamauchi et al. reported the reduction of(2-oxocyclohexyl)acetate with baker's yeast to provide anenantiomerically pure starting materials, which were then used toprepare 2,6-disubstituted piperidines (Biosci. Biotechnol. Biochem.68(3):676-84, 2004). Further examples of synthetic routes to2,6-disubstituted piperidine alkaloids as disclosed herein can be foundin, for example, Felpin et al. Curr. Org. Syn. 1:83-109, 2004; Comins etal. J. Org. Chem. 56:2506, 1991; and Wang et al. J. Org. Chem.70:1897-1900, 2005. In yet another example, methods for preparingSolenopsin derivatives and analogous piperidine alkaloids are disclosedin U.S. Pat. No. 6,369,078. Each of these references is incorporated byreference herein in its entirety at least for its teachings ofpiperidine alkaloids and methods for preparing them.

Methods for the chemical synthesis of piperidine alkaloids such assolenopsins are disclosed in U.S. Patent Publication 2005/0038071, whichis incorporated herein by reference in its entirety for the teaching ofthese compositions and methods. An efficient flexible chemistry can beused to synthesize Solenopsin A and analogues from Solenopsin A. Variousmethods can be used and adapted for a number of analogues disclosedherein. See, for example, Comins, D. L., et al. (J. Org. Chem. 1991, 56,2506), which is incorporated herein by reference for these teachings.For example, 4-chloropyridine can undergo introduction of an R group atthe 2-position of the pyridine ring using alkylmagnesium bromide in THFat −78° C. followed by treatment with phenylchloroformate to provide therespective dihydropyridine derivative. The dihydropyridine derivativecan then be converted into the corresponding N-Boc (Boc is a tertiarybutyl carbonate group) derivative using potassium t-butoxide intetrahydrofuran and −42° C. A methyl (or other alkyl group) can beintroduced into the 6 position of the dihydropyridine ring by utilizinga first step of n-butyllithium (n-BuLi) in THF at −78° C. followed byintroduction of the methyl(alkyl) group at the 6 position of thedihydropyridine compound utilizing methyliodide to form the dialkylsubstituted chlorine substituted dihydropyridine derivative. The dialkylsubstituted chorine substituted dihydropyridine derivative can then besubjected to a hydrogenation procedure (hydrogen, palladium/carboncatalyst in methanol) to remove the chlorine group at the 4 position,which derivative is further hydrogenated using NABH₃/TFA in methylenechloride to provide the dialkyl substituted N-boc piperidine derivative.The boc group can be readily removed using 15% trifluoracetic acid inmethylene chloride to afford the dialkyl substituted piperidinederivative. Salt formation can readily occur using the appropriate acidto acidify the basic nitrogen.

In an alternative chemical synthetic method, a more efficient route tothe dialkyl substituted piperidine analogues can be used. This route canallow the facile introduction of a double bond in the side chain of the2 position of the piperidine ring. The method can follow the chemicalmethods reported by Beak, et al. (Beak, P.; Lee, W. K. J. Org. Chem.1993, 58, 1109; Tetrahedron Lett. 1989, 30, 1197), which areincorporated herein by reference for the teaching of this syntheticmethod. This method demonstrated a regioselective and diastereoselectivemethod for a lithiation-substitution at a methylene group.

N-boc piperidine can be subjected to sec-butyllithium (sec-BuLi) at −78°C. followed by dimethylsulfate to provide the methyl substituted N-Bocpiperidine analog. The N-Boc piperidine analog prepared above can thenbe subjected to sec-BuLi at −78° C. followed by dimethyl formamide toproduce the formyl piperidine derivative which can be further reactedusing a Wittig procedure to produce longer chain alkylated products(saturated or unsaturated). If one desires an unsaturated side chain,the Wittig reaction can afford such a substituent directly, followed byremoval of the Boc group using procedures. Salt formation may alsoreadily occur, using standard methods available in the art.

Alternatively, to provide the alkyl (saturated) side chains, the Wittigproduct can be reduced using hydrogen/Pd/C to provide the fullysaturated side chain. The Boc group can be readily removed using thepreviously described method, followed by salt formation.

The above-described methods of chemical synthesis can be readily adaptedby those of ordinary skill to substitute different side chains at the 2and 6 position of the piperidine ring to produce the disclosedcompounds. These methods can be readily adapted to produce a largenumber of side chains for the disclosed compounds.

4. Method of Administering

The compositions may be administered orally, parenterally (e.g.,intravenously), by intracranial injection, by intramuscular injection,by intraperitoneal injection, transdermally, extracorporeally, topicallyor the like, including topical intranasal administration oradministration by inhalant. Administration of the compositions byinhalant can be through the nose or mouth via delivery by a spraying ordroplet mechanism. Delivery can also be directly to any area of therespiratory system (e.g., lungs) via intubation. The exact amount of thecompositions required will vary from subject to subject, depending onthe species, age, weight and general condition of the subject, theseverity of the disorder being treated, the particular composition used,its mode of administration and the like. Thus, it is not possible tospecify an exact amount for every composition. However, an appropriateamount can be determined by one of ordinary skill in the art using onlyroutine experimentation given the teachings herein.

Parenteral administration of the composition, if used, is generallycharacterized by injection. Injectables can be prepared in conventionalforms, either as liquid solutions or suspensions, solid forms suitablefor solution of suspension in liquid prior to injection, or asemulsions. Another approach for parenteral administration involves useof a slow release or sustained release system such that a constantdosage is maintained. See, e.g., U.S. Pat. No. 3,610,795, which isincorporated by reference herein.

The materials may be in solution, suspension (for example, incorporatedinto microparticles, liposomes, or cells). These may be targeted to aparticular cell type via antibodies, receptors, or receptor ligands. Ingeneral, receptors are involved in pathways of endocytosis, eitherconstitutive or ligand induced. These receptors cluster inclathrin-coated pits, enter the cell via clathrin-coated vesicles, passthrough an acidified endosome in which the receptors are sorted, andthen either recycle to the cell surface, become stored intracellularly,or are degraded in lysosomes. The internalization pathways serve avariety of functions, such as nutrient uptake, removal of activatedproteins, clearance of macromolecules, opportunistic entry of virusesand toxins, dissociation and degradation of ligand, and receptor-levelregulation. Many receptors follow more than one intracellular pathway,depending on the cell type, receptor concentration, type of ligand,ligand valency, and ligand concentration. Molecular and cellularmechanisms of receptor-mediated endocytosis has been reviewed (Brown andGreene, DNA and Cell Biology 10:6, 399-409, 1991).

5. Doses

Effective dosages and schedules for administering the compositions maybe determined empirically, and making such determinations is within theskill in the art. The dosage ranges for the administration of thecompositions are those large enough to produce the desired effect inwhich the symptoms disorder are effected. The dosage should not be solarge as to cause adverse side effects, such as unwantedcross-reactions, anaphylactic reactions, and the like. Generally, thedosage will vary with the age, condition, sex and extent of the diseasein the patient, route of administration, or whether other drugs areincluded in the regimen, and can be determined by one of skill in theart. The dosage can be adjusted by the individual physician in the eventof any counterindications. Dosage can vary, and can be administered inone or more dose administrations daily, for one or several days.Guidance can be found in the literature for appropriate dosages forgiven classes of pharmaceutical products. For example, guidance inselecting appropriate doses for antibodies can be found in theliterature on therapeutic uses of antibodies, e.g., Handbook ofMonoclonal Antibodies, Ferrone et al., eds., Noges Publications, ParkRidge, N.J., (1985) ch. 22 and pp. 303-357; Smith et al., Antibodies inHuman Diagnosis and Therapy, Haber et al., eds., Raven Press, New York(1977) pp. 365-389. A typical daily dosage of the antibody used alonemight range from about 1 μg/kg to up to 100 mg/kg of body weight or moreper day, depending on the factors mentioned above.

Following administration of a disclosed composition for treating,inhibiting, or preventing a neurological disease, the efficacy of thetherapeutic can be assessed in various ways well known to the skilledpractitioner. For instance, one of ordinary skill in the art willunderstand that a composition disclosed herein is efficacious intreating or inhibiting a neurological disease in a subject by evaluatingcognitive function.

D. SPECIFIC EMBODIMENTS

Provided is a method of treating or preventing neurodegenerative diseasein a subject comprising identifying a subject at risk forneurodegenerative disease and administering to the subject atherapeutically effective amount of a composition comprising apiperidine alkaloid. The composition can comprise an extract of fire ant(Solenopsis invicta) venom. The piperidine alkaloid can comprises theformula

wherein R is n-alkyl-piperidine.

The piperidine alkaloid can comprises 2-methyl-6-alkyl-piperidine. Thepiperidine alkaloid can be Solenopsin A. The method piperidine alkaloidcan be selected from the group consisting ofcis-2-methyl-6-undecylpiperidine, trans-2-methyl-6-undecylpiperidine,cis-2-methyl-6-tridecylpiperidine, trans-2-methyl-6-tridecylpiperidine,cis-2-methyl-6-pentadecylpiperidine,trans-2-methyl-6-pentadecylpiperidine,cis-2-methyl-6-(cis-4-tridecen-1-yl)piperidine,trans-2-methyl-6-(cis-4-tridecen-1-yl)piperidine,cis-2-methyl-6-(cis-4-pentadecen-1-yl)piperidine,trans-2-methyl-6-(cis-4-pentadecen-1-yl)piperidine. The piperidinealkaloid can be 2-methyl-6-undecylpiperidine.

In one aspect, the subject has been diagnosed with Alzheimer's disease.The composition can be administered to the subject orally, systemically,or intracranially.

Also provided is a method of treating a subject at risk for Alzheimer'sdisease comprising administering to the subject a composition comprisinga piperidine alkaloid.

E. EXAMPLES 1. Example 1 Animal Study

Fire ant venom (FAV) was extracted from S. invicta from Charleston, S.C.and transported in a Teflon beaker to prevent the ants from escaping.The ants were placed (n=100) in 30 ml vials containing hexane (5 ml).The vials were then sealed and the contents analyzed for the presence ofFAV alkaloids after 24 h. The ants released their venom upon contactwith the solvent.

Fire ant venom (FAV) extract was injected into aged rats (20 months) atseveral different doses (1, 10, and 100 ng/kg (i.p., daily). Animalswere subjected to behavioral analysis using a Morris Water Maze andopen-field testing after 7 days. Animals demonstrated a dose-dependentimprovement in age-related deficits compared to vehicle injectedanimals. In addition, no adverse effects were seen in the animals at thedoses given.

2. Example 2 Clinical Study

Mr. D., who at the time was an 86 year old, widowed, white male born inthe U.S. Clinical Observations included 25 lb. weight loss, listless,restless, depressed, disorientation, and anxiety. Patient exhibited asignificant decrease in cognitive functions, particularly in the areasof orientation (time, place, person), recent memory, new learningability, digit span, information and vocabulary, calculating ability,and abstract thinking characteristic of mild to moderate Alzheimer'sdisease.

After serendipitous exposure to FAV (20-30 bites, 20-30 units/1 μl)within a 48-72 hour window patient exhibited marked improvement incognitive baseline, especially in the aforementioned areas; and infollowing this patient during the last year (with no maintenancedosing), although patient has demonstrated some cognitive regression hestill remains markedly improved in cognitive abilities. In addition, allsigns of depression are gone; he has gained 27 lbs. during that one-yeartime frame, goes to the gym four times per week, and has resumed anactive social lifestyle among his elderly peer group. Friends and familyhave remarked about the remarkable turnaround in his life.

Patient presents a unique pristine pharmacological history as the onlymedication he was and is presently taking is 81 mg of aspirin per day.

What is claimed is:
 1. A method of treating a human subject diagnosedwith Alzheimer's disease, comprising administering to the human subjecta therapeutically effective amount of a composition comprising apiperidine alkaloid, wherein the piperidine alkaloid comprises theformula

wherein R¹ is a substituted or unsubstituted aliphatic group having from10 to 20 carbon atoms.
 2. The method of claim 1, wherein the compositioncomprises an extract of fire ant (Solenopsis invicta) venom.
 3. Themethod of claim 1, wherein the piperidine alkaloid is Solenopsin A. 4.The method of claim 1, wherein the piperidine alkaloid is selected fromthe group consisting of trans-2-methyl-6-undecylpiperidine,trans-2-methyl-6-tridecylpiperidine, andtrans-2-methyl-6-pentadecylpiperidine.
 5. The method of claim 4, whereinthe piperidine alkaloid is trans-2-methyl-6-undecylpiperidine.
 6. Themethod of claim 1, wherein the composition is administered to thesubject orally, systemically, or intracranially.